Journal: PLoS ONE

Article Title: Generation of Virus-Free Induced Pluripotent Stem Cell Clones on a Synthetic Matrix via a Single Cell Subcloning in the Naïve State

doi: 10.1371/journal.pone.0038389

Figure Lengend Snippet: (A) Phase contrast light microscopic observation of cells during reprogramming and recloning. Images captured on a Pronectin F-coated dish prior to colony picking on days four, nine, 13, 17 and 24 (upper panels). Note that human ES cell-like colonies emerged within a cobblestone like morphology. (B) Efficiency of generating reprogrammed cells on various coating materials and the number of colonies characterized. All the experiments used 1×10 4 CD34 + CBCs, SeV TS vectors at 20 M.O.I. and ReproFF medium. Three independent experiments for Matrigel, Pronectin F and two independent experiments for fibronectin and laminin (Laminin-extracts), Pro nectin L we re performed. (C) Frequency of generating human ES cell-like colonies in various culture media. Five thousand CD34 + CBCs were infected with 20 M.O.I. of SeV carrying four factors and cultured in ReproFF, ReproFF2, mTeSR1, or E8 medium on Pronectin F-coated dishes to reprogram CD34+ CBCs. D: Endogenous gene expression of Klf4, c-Myc, Oct3/4, and Sox2 in feeder like-cells (lane 1) and first pick up of a human ES cell like-colony (lane 2).

Article Snippet: The culture dish (BD Life Science) was covered completely with fibronectin (Sigma) or laminin extracts (ReproCELL) and left overnight at room temperature.

Techniques: Infection, Cell Culture, Expressing